Method of treating cancer with a metallic nanoparticle composition

ABSTRACT

The invention is directed to a composition of metal particles and methods of manufacturing and using the composition in the treatment of microbial infections and cancer. The particles can be nanoparticles having coupled thereto at least one of a surfactant, an antibiotic, and a drug. The particles of the invention achieve enhanced stability, enhanced cytotoxicity, and enhanced antimicrobial activity through novel combinations of metals, surfactants, antibiotics, and drugs.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.17/086,374, filed Oct. 31, 2020, which claims the benefit of U.S.Provisional Application No. 62/929,241 filed Nov. 1, 2019, and U.S.Provisional Application No. 62/979,562 filed Feb. 21, 2020, the entirecontents of the forgoing applications are incorporated herein byreference for all purposes.

FIELD OF INVENTION

The invention generally relates to nanotechnology. More particularly,the invention relates to a cytotoxic, antimicrobial nanoparticlecomposition and methods of its use and manufacture in therapeutic,antiseptic, and other applications.

BACKGROUND

Spread of harmful bacteria, fungi, viral infections and cancer are amongthe utmost challenges humanity faces on daily basis. More and more drug-and multidrug-resistant bacterial pathogens are identified by thescientific community every year. In 2014, the World Health Organization(WHO) warned that humanity is approaching the post-antibiotic era; aworld in which antibiotics will no longer be effective, and even minutecontaminations will be life threatening.

Small silver nanoparticles, the size of proteins, are known to corrodeand dissolve in vivo yielding Ag+ ions that induce reactive oxygenspecies (ROS) and interfere with respiration mechanisms of bacteriaresulting in a very broad-spectrum antibiotic. Silver nanoparticles havereceived significant attention as powerful antimicrobial and neoplasticagents resulting from their high surface-to-volume ratios and theirunique physical and chemical properties [11-13]. Silver nanoparticleshave been demonstrated to act as effective antibacterial agents againstvarious multidrug-resistant bacteria like MRSA and methicillin-resistantS. epidermidis bacteria [14]. However, silver nanoparticles haverelatively low stability in air and their size and shape are difficultto control in an aqueous environment. Moreover, silver nanoparticlepreparations require concentrated solutions to be effective [15].

Chemotherapeutic drugs containing platinum are first- and second-linetherapy for many types of cancer. Cisplatin, for example, is used intesticular, ovarian, lung, and head and neck cancer [16]. Cisplatin'sclinical use, however, is aggravated with systemic toxicity, primarilyto the kidneys [17]. A series of improved analogues have been developedsince Cisplatin was approved by the FDA in 1978. Still, the mainlimitation to the clinical usefulness of Cisplatin (and itsmodifications) as anticancer drugs is the high incidence ofchemoresistance [18]. Generally, cancer therapies based on a single drugare infective due to the complex microenvironment of cancer cells anddrug resistance mechanisms [19]. Also, the use of chemotherapeutic drugsincreases the risk of infections. Thus, there is a need for agents thathave both antibacterial and anticancer effects against drug-resistantcancer cells.

What is needed in the art therefor is an improved cytotoxic,antimicrobial nanoparticle therapeutic with improved stability,consistency, and greater potency against drug resistant infections andcancers.

SUMMARY OF THE INVENTION

The invention provides a nanoparticle composition having synergisticallyenhanced stability and efficacy in the treatment of drug-resistantinfections and tumors. The invention achieves this and other objectivesby providing unique combinations of metals, surfactants, antibiotics,and drugs.

It is therefore an object of the invention to provide a compositioncomprising (i) particles of at least one metal, and (ii) at least onesurfactant.

In some aspects, the metal is a noble metal.

In some aspects, the metal is selected from the group consisting ofsilver, gold, platinum, palladium, osmium, iridium, rhodium, ruthenium,and combinations thereof.

In some aspects, the particles include monometallic particles,polymetallic particles, or a combination thereof.

In some aspects, the particles include bimetallic particles.

In some aspects, the bimetallic particles are silver and gold bimetallicparticles, silver and platinum bimetallic particles, or a combinationthereof.

In some aspects, the composition comprises about 99.67% molar silver,about 99.50% molar silver, about 97.50% molar silver, about 70% molarsilver, or about 50% molar silver.

In some aspects, the particles are polymetallic particles having a coreof a first one or more metals, and a shell of a second one or moremetals.

In some aspects, the particles are bimetallic particles having a core ofgold or platinum and a shell of silver.

In some aspects, the particles are polymetallic particles having astructure selected from an alloy, a mixed alloy, a subcluster with twoor more interfaces, and combinations thereof.

In some aspects, the subclusters are segregated subclusters, mixedsubclusters, or a combination thereof.

In some aspects, the alloy is an intermetallic alloy.

In some aspects, the composition comprises silver monometallicparticles, gold monometallic particles, platinum monometallic particles,or combinations thereof.

In some aspects, the particles are nanoparticles.

In some aspects, the nanoparticles have a mean diameter that is betweenabout 5 nanometers and about 30 nanometers, or between about 5nanometers and about 400 nanometers.

In some aspects, the particles have a shape that is substantiallyspherical, substantially oval, or substantially cuboidal.

In some aspects, the surfactant is selected from one or more of acationic surfactant, an anionic surfactant, an amphoteric surfactant,and a charge-neutral surfactant.

In some aspects, the surfactant is selected from one or more ofbenzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride, acetyltrimethylammonium salt, a benzalkonium salt,didecyldimethylammonium chloride, octenidine dihydrochloride, dimethylbenzyl ammonium chloride, a polyhexamethylene biguanide, apolyhexamethylene guanidine, a polyhexamethylene biguanide salt, apolyhexamethylene guanidine salt, sodium lauryl ether sulfate, andsodium cocaminopropionate.

In some aspects, the surfactant is coupled to the surface of theparticles.

In some aspects, the composition further comprises an antibiotic.

In some aspects, the antibiotic is at least one of tetracycline andvancomycin.

In some aspects, the antibiotic is coupled to the surface of theparticles, coupled to the surfactant, or a combination thereof.

In some aspects, the composition further comprises a drug.

In some aspects, the drug is chemotherapeutic cancer drug.

In some aspects, the drug is one or more of doxorubicin, glucosamine,and metformin.

In some aspects, at least a portion of the drug molecules are coupled tothe surface of the particles, coupled to the surfactant, coupled to theantibiotic, or combinations thereof.

In some aspects, the composition further comprises at least one of acarrier and an excipient, wherein at least a portion of one or more ofthe surfactant, the antibiotic, and the drug is dissolved in orsuspended in the carrier and excipient.

In some aspects, the composition is a colloid.

In some aspects, the composition is in a form selected from a liquid,gel, sol, and foam.

In some aspects, the composition is in an administration form selectedfrom a pill, capsule, tablet, microbead, injection, infusion, andsuppository.

In some aspects, the composition is in contact with a bandage or wounddressing.

In some aspects, the composition is in contact with a textile, such as abed sheet, blanket, pillow, pillow case, seat cover, table cover, doormat, gauze, surgical mask, surgical gown, patient gown, menstrual pad,or tampon.

In some aspects, the invention provides a method of preventing orinhibiting microbial growth on or in a material, comprising contactingthe material with the composition.

In some aspects, the invention provides a method of treating a microbialinfection, comprising administering the composition to a patient in needthereof.

In some aspects, the patient has at least one of a bacterial infection,viral infection, and a fungal infection.

In some aspects, the invention provides a method of treating cancercomprising administering the composition to a patient in need thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an electron micrograph of silver nanoparticles from Sample1-1.

FIG. 2 is an electron micrograph of silver-gold bimetallic nanoparticlesfrom Sample 1-3.

FIG. 3 shows the absorbance spectra of the nanoparticles of Example 1,wherein Sample 1-1 is represented as “Ag,” Sample 1-2 is represented as“1,” Sample 1-3 is represented as “2,” and Sample 1-4 is represented as“3.”

FIG. 4 shows the size distribution of the silver monometallicnanoparticles of Sample 1-1.

FIG. 5 shows the size distribution of the silver-gold bimetallicnanoparticles of Sample 1-3.

FIG. 6 is a schematic diagram of bimetallic nanoparticles having a goldcore and a silver shell with a surfactant on the surface of the shell.

FIG. 7 shows the absorbance spectra of the nanoparticles of Example 6,wherein Sample 4-1 is represented as “2,” Sample 4-2 is represented as“1,” and Sample 4-3 is represented as “3.”

FIGS. 8A-8C show nanoparticle size distribution of the nanoparticles ofExample 7, wherein FIG. 8A shows size distribution for Sample 4-1, FIG.8B shows size distribution for Sample 4-2, and FIG. 8C shows sizedistribution for Sample 4-3.

FIG. 9 shows the absorbance spectra of the nanoparticles of Examples 10and 11, wherein Sample 6-1 is represented as “3,” Sample 6-2 isrepresented as “1,” and Sample 7-1 is represented as “2.”

FIGS. 10A-10C show TEM images of the nanoparticles of Examples 10 and11, wherein FIG. 10A represents Sample 6-1, FIG. 10B represents Sample6-2, and FIG. 10C represents Sample 7-1.

FIGS. 11A-11C show the size distribution of the nanoparticles ofExamples 10 and 11, wherein FIG. 11A represents Sample 6-1, FIG. 11Brepresents Sample 6-2, and FIG. 11C represents Sample 7-2.

DEFINITIONS

As used herein, the term “about” refers to the quantity, level, value,number, frequency, percentage, dimension, size, amount, weight or lengththat is referenced, or that varies (plus or minus) by as much as 30%,25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of thereferenced quantity, level, value, number, frequency, percentage,dimension, size, amount, weight or length.

The terms “a” and “an,” when used in conjunction with the word“comprising” in the claims or specification, denotes one or more, unlessspecifically noted.

The terms “is” and “are,” when used in conjunction with a group ofdifferent possibilities for a limitation or element in the claims orspecification, indicates the group includes, but is not limited to, thelisted possibilities unless expressly specified otherwise.

As used herein, the term “metal” refers to any transition metal,including, without limitation, the noble metals. The metal can be anelemental metal, metal ion, or metal salt.

As used herein, the term “monometallic” refers to a single metal.

As used herein, the term “polymetallic” refers to a combination of two,three, four, five, or more metals.

As used herein, the term “bimetallic” refers to a combination of justtwo metals.

As used herein, the term “surfactant” refers to a compound that lowersthe surface tension between two liquids, between a gas and a liquid, orbetween a liquid and a solid.

Where this disclosure refers to two or more possible limitations orelements in the alternative, such as by use of the term “or,” it iscontemplated that one or more of the listed limitations or elements canbe specifically excluded from the embodiment that is being described.

DETAILED DESCRIPTION

The inventors surprisingly discovered that the stability of metalparticles and their cytotoxic, antimicrobial effects, can besynergistically increased through unique combinations of the metals thatare used, as well as their combination with one or more of a surfactant,antibiotic, and drug.

In some embodiments, the invention provides a composition comprisingparticles of at least one transition metal and one or more surfactants.In some non-limiting embodiments, the transition metal is a noble metal.The noble metal can be one or more of silver, gold, platinum, palladium,osmium, iridium, rhodium, and ruthenium. In other non-limitingembodiments, the metal is one or more of silver, gold, platinum,palladium, osmium, iridium, rhodium, ruthenium, copper, chromium, andiron. The particles can be monometallic, bimetallic, or polymetallicwith three or more metals, or mixtures of such particles. Thecomposition can comprise a combination of monometallic particles ofdifferent metals. For example, the composition can comprise acombination of silver particles and gold particles. Similarly, thecomposition can comprise monometallic particles in combination withpolymetallic particles. For example, the composition can comprisemonometallic silver particles, monometallic gold particles, andbimetallic silver and gold particles. In a non-limiting embodiment, thecomposition comprises bimetallic particles wherein the particles aregold and silver, silver and platinum, gold and platinum, or combinationsthereof.

When the composition comprises particles of more than one metal, whetheras polymetallic and/or monometallic particles, the composition can beformulated to contain a specific molar percentage of the metals. Themolar percentage of a metal in the composition can be determined bydividing the number of mols of the metal being quantified, by the totalnumber of mols of all metals that are present in the composition, itbeing understood that the molar percentages are quantified based on theamounts elemental metal and metal ions present in the composition. Amolar percentage can be expressed as one or a combination of metals. Ina non-limiting embodiment, the composition can be formulated to containa first one or more metals in a molar percentage of about 1%, about 5%,about 10%, about 20%, about 30%, about 40%, or about 50%, with thesecond one or more metals making up the remainder of the molarpercentage. For example, and by no way of limitation, the compositioncan be formulated to contain about 1% molar gold and about 99% molarsilver, about 5% molar gold and about 95% molar silver, about 10% molargold and about 90% molar silver, about 20% molar gold and about 80%molar silver, about 30% molar gold and about 70% molar silver, about 40%molar gold and about 60% molar silver, or about 50% molar gold and about50% molar silver. The molar percentages disclosed herein can refer tothe molar percentage of the metals in the form of particles, with orwithout any metal ions that may be present in the composition as asolute or salt.

Particles of the composition can have a shape that is substantiallyspherical, substantially cuboidal, or substantially oval. The particlesof the composition can have the same shape, or a mixture of shapes. Forexample, the composition can comprise a first portion of particles thatis substantially spherical, and a second portion of particles that issubstantially oval. As used herein, the term “substantially” can referto the shape that is referenced, or that resembles the shape that isreferenced. The particles can also assume a variety of structures. Thestructure of the particles can be a core and shell structure, an alloy(e.g. mixed alloy), or a subcluster. The particles of the compositioncan have the same shape, or comprise a mixture of particles of differentshapes. The core and shell structure can comprise a core of a first oneor more metals that is surrounded by one or more shells made from asecond one or more metals. Alloy particles for use with the inventioncan be an intermetallic alloy. Subclusters for use with the inventioncan be segregated subclusters, mixed subclusters, or subclusters withone or more interfaces. The composition can comprise one of these typesof subclusters, or a mixture of these subclusters. The particles canhave a structure as disclosed in the following publication, the entirecontents of which are incorporated herein for all purposes: Srinoi,Appl. Sci. 2018, 8, 1106. Core shell structures can be a core of a firstone or more noble metals and at least one shell of a second one or morenoble metals. FIG. 6 shows a non-limiting embodiment of bimetallicparticles that have gold core 601 surrounded by a silver shell 602 withsurfactant molecules 603 coupled to silver shell 602.

In some embodiments, at least a portion of the particles in thecomposition are nanoparticles. The nanoparticles can have a mean sizethat ranges between about 2 nanometers and about 15 nanometers, betweenabout 5 nanometers and about 30 nanometers, or between about 2nanometers and about 400 nanometers. The nanoparticles can have a meansize of up to about 15 nanometers, up to about 30 nanometers, or up toabout 400 nanometers.

In some embodiments, the composition comprises one or more surfactants.Surfactants can be present in the composition in an amount that isbetween about 0.001 w/w % and about 0.10 w/w %, or between about 0.00001w/w % and about 5-10 w/w %. Surfactants in the composition can bepresent in an amount that is between about 0.1 ppm and about 10,000 ppm,or between about 10 ppm and about 1,000 ppm. The surfactant can be oneor more surfactants selected from a cationic surfactant, an anionicsurfactant, an amphoteric surfactant, and a charge-neutral surfactant.Non-limiting examples of suitable surfactants include:benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®); cetyltrimethylammonium salts; benzalkonium salts (such asbenzalkonium chloride); didecyldimethylammonium chloride; octenidinedihydrochloride; dimethyl benzyl ammonium chloride; polyhexamethyleneguanidine; polyhexamethylene guanidine salt; polyhexamethylenebiguanide; polyhexamethylene biguanide salt (such as polyhexamethylenebiguanide chloride); sodium lauryl ether sulfate; and sodiumcocaminopropionate. The composition can comprise just one surfactant ora combination of surfactants. Without being limited to any particulartheory, mechanism, or effect, the combination of the metal particleswith a surfactant inhibits oxidation of the metals in the particlesthereby increasing their stability and preserving their efficacy.Moreover, and without being limited to any particular theory mechanism,or effect, combining the particles with a surfactant increaseshomogeneity of the shape and size of the particles, inhibits particleaggregation, stabilizes the net charge of the particles, and improvestheir dispersity when suspended in a medium.

The surfactant can be coupled to the surface of the particles. Thesurfactant can be coupled to the surface of the particles by at leastone of ionic bonding, hydrogen bonding, dipole-dipole interaction,covalent bonding, and donor-acceptor interaction.

In some embodiments, the composition further comprises at least one ofan antibiotic and a drug. The antibiotic and drug can be present in anamount that is between about 0.000001 w/w % and about 1-5 w/w %, orbetween about 0.0001 and about 0.01 w/w %. In some non-limitingembodiments, the ratio of the surfactant to the antibiotics and/or drugsis between about 2:1 and about 10:1. The ratio of the surfactant to theantibiotics and/or drugs can about 2; 1, about 3:1, about 4:1, about5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1. Theantibiotic and/or drugs can be present in an amount that is betweenabout 0.01 ppm and about 2 ppm, or between about 0.1 ppm and about 0.5ppm. The antibiotic can be, without limitation, one or more oftetracycline, vancomycin, β-lactams (including ampicillin, amoxicillin,penicillin, methicillin etc.), quinolones (enoxacin, ofloxacin,norfloxacin, ciprofloxacin etc.), aminoglicosides (kanamycin A,amikacin, tobramycin, etc.), tetracycline type (tetracycline,chlortetracycline, oxytetracycline, etc.), glycopeptide antibiotics(vancomycin, teicoplanin, telavancin, etc.), anthracycline type(doxorubicin, daunorubicin, epirubicin, idarubicin etc.) and otherclasses of antibiotics, bioactive amines, and aromatic compounds. Insome aspects, the drug is a cancer drug Non-limiting examples of drugsfor use with the invention include one or more of doxorubicin,glucosamine, and metformin. The antibiotic and/or drug can be in contactwith at least one of the surface of the particles and the surfactant.The antibiotic and/or drug can be coupled to the surface of theparticles, the surfactant, and/or one another by ionic bonding, hydrogenbonding, dipole-dipole interaction, covalent bonding, donor-acceptorinteraction, or combinations thereof. In some embodiments, the drug isin present in the composition as a solution or suspension.

In at least one embodiment, the composition comprises a colloid whereinthe particles are suspended in a medium, such as a liquid, gel, or sol.The surface of the particles in the colloid can be in contact with orcoupled to one or more surfactants, one or more antibiotics, one or moredrugs, or a combination thereof. At least a portion of at least one ofthe surfactant, antibiotic, and drug can be free of contact with thesurface of the particles and independently suspended or dissolved withinthe medium. Without being limited to any particular theory, mechanism oreffect, contacting or coupling the surfactant with the surface of theparticles increases colloidal stability of the composition. The colloidcan contain a dispersed phase comprising the particles in contact withor coupled to at least one of a surfactant, antibiotic, and drug, and acontinuous phase comprising the medium.

In one non-limiting embodiment, the invention provides achemotherapeutic composition of polymetallic nanoparticles having a coreof a first one or more noble metals, at least one shell of a second oneor more noble metals substantially surrounding the core, and at leastone surfactant on the surface of the outermost shell of thenanoparticles.

In at least one embodiment, the composition is a stable suspension ofcolloidal silver-gold stabilized with a surfactant. The silver contentin the composition can be between about 0.00001 w/w % and about 5-10 w/w%. The gold content in the composition can be between about 0.00001 w/w% and about 5-10 w/w %. The total content of the surfactants in thecomposition can be between about 0.00001 w/w % and about 5-10 w/w %. Inat least some embodiments, the composition comprises between about 0.001w/w % and about 0.10 w/w % of least one metal, and between about 0.001w/w % and about 0.10 w/w % of at least one surfactant.

Metal particles can be produced by reduction methods known in the art.The following reducing agents can be used in the manufacture metalparticles: sodium borohydride, citric acid, salts of citric acid(citrate), ascorbic acid, glucose, or combinations thereof.

In at least some embodiments, the composition is formulated to beadministered to a patient. The composition can be formulated with apharmaceutically acceptable excipient, pharmaceutically acceptablecarrier, or combination thereof. Suitable excipients and carriers foruse with the invention include, but are not limited to, those disclosedin the following references, the entire contents of which areincorporated herein by reference for all purposes: The Science andPractice of Pharmacy, 19^(th) Ed (Easton, Pa.: Mack Publishing Company,1995); Hoover, John E., Remington's Pharmaceutical Sciences, (Easton,Pa.: Mack Publishing Co 1975); Liberman, H. A. and Lachman, L., Eds.,Pharmaceutical Dosage Forms (New York, N.Y.: Marcel Decker 1980); andPharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed(Lippincott Williams & Wilkins 1999). The composition can assume adosage form suitable for administration to a patient, including, but notlimited to, a powder, liquid, pill, tablet, pellet, capsule, thin film,solution, spray, syrup, linctus, lozenge, pastille, chewing gum, paste,vapor, suspension, emulsion, ointment, cream, lotion, liniment, gel,drop, topical patch, buccal patch, bead, beadlet, gummy, gel, sol,injection, or combinations thereof.

In at least one aspect, the invention provides a drug delivery platformwherein nanoparticles comprising one or more metals and a surfactantenhance the delivery drugs and antibiotics into cells and microbes.Without being limited to any particular theory or mechanism, thesurfactant on the surface of the nanoparticle facilitates permeation ofcell membranes such that nanoparticles bearing drugs and antibiotics arepermitted to enter cells and microbes with greater efficiency therebyincreasing the intracellular concentration of the therapeutic. In thecase of cells, the acidic environment of intracellular lysosomesliberate metal ions from the nanoparticles to permit the ions to exert acytotoxic effect.

In some aspects, the invention provides a method of antisepsis,comprising contacting a microbe with a composition as disclosed herein.As used herein, the terms “microbe,” “microbial,” and the like refer, tobacteria, fungi, algae, parasite, and viruses. The microbe can be on orin a material where microbial antisepsis is desired. In other aspects,the invention provides a method for preventing or inhibiting microbialgrowth on or in a material, comprising contacting the material with acomposition as disclosed herein. In both methods, the material can be asurface, such as the surface of a table or shelve, flooring, wall, amat, a textile, a medical or surgical device, an appliance, a storagedevice, a container, a bandage or a wound dressing, for example. Thematerial can be water, such swimming pool water, water in agriculturaland aquaculture ponds applications, river water, lake water, reservoirwater, or water in water treatment facility applications. Thecomposition can be contacted with water to prevent or inhibit the growthof algae in the water.

In at least one embodiment, the invention provides a method for treatinga microbial infection, comprising administering to a patient in needthereof a composition as disclosed herein. In other embodiments, theinvention provides a method of treating cancer, comprising administeringto a patient in need thereof a composition as disclosed herein. Thecancer can be gliobastoma or ovarian cancer. In both methods oftreatment, the patient can be a human. The patient can be an animal,including, without limitation, dogs, cats, cattle, horses, sheep, goats,chicken, turkeys, rats, mice, and primates. In both methods oftreatment, the composition can be administered systemically or locally.The composition can be administered orally, parenterally, or acombination thereof. The composition can be administered intravenously,intraarterially, sublingually, intravaginally, rectally, topically,sub-dermally, intramuscularly, intranasally, intraocularly,intra-aurally, or combinations thereof.

The invention is demonstrated by the following examples, it beingunderstood that the examples are merely illustrative and are notintended to limit the scope of the present invention as one skilled inthe art will appreciate that variations may be possible based on theteachings of the specification herein.

EXAMPLES

The following examples exemplify some, but not necessarily all, of thevarious embodiments of the invention. The following examples areprovided purely for illustrative purposes and do not in any way limitthe scope of the present invention, it being understood that the scopeof the invention is set forth in the claims and their equivalents.

Example 1—Nanoparticle Preparation—Bimetallic Nanoparticle CompositionHaving Miramistin®

A composition of silver and gold bimetallic nanoparticles with(Miramistin®) was prepared as follows: aqueous solutions of silver andgold salts were added to a solution of at least one surfactant undervigorous stirring, then a reducing agent was added. The reaction wascarried out in an inert atmosphere of nitrogen or argon. Silver nitrateor silver acetate can be used as the silver salt, and hydrogentetrachloroaurate as a source of gold.

The following procedure was used for the preparation of Sample 1-2.Samples 1-3 and 1-4 were prepared according to the same procedure, withadjustments made to the amounts of the metal-bearing components toachieve the molar percentages of the respective samples. Distilled waterwas repeatedly distilled in the atmosphere of nitrogen gas to achievede-oxygenation. The de-oxygenized water was used for all furtherpreparations. Aqueous solutions of 20 mg silver nitrate and 0.15 mghydrogen tetrachloroaurate in 10 mL water were added dropwise tovigorously stirred 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) in water. The resulting mixture was stirred for 15 min,followed by dropwise addition of aqueous 0.01 g sodium borohydride in100 mL water. After addition of all the reagents, the reaction mixturewas stirred for another hour.

The monometallic silver nanoparticle composition of Sample 1-1 (havingMiramistin® but lacking metallic gold) was obtained using a processknown in the art and was used as a control.

Table 1 shows the silver-gold molar percentages of the preparations thatwere obtained.

TABLE 1 Sample No. Au molar % in Sample Sample 1-1 0 Sample 1-2 0.33Sample 1-3 0.50 Sample 1-4 2.50

Example 2—Nanoparticle Characterization—Bimetallic NanoparticleComposition Having Miramistin®

UV-V is absorption spectra were measured to confirm nanoparticleformation. FIG. 3 shows the absorption spectra of the nanoparticles inthe resulting compositions. Incorporation of gold affects the dispersityof nanoparticles in the final product, the absorption spectra ofbimetallic particles displayed more intense, narrow and symmetricalabsorption bands. At 0.5% gold molar content, the nanoparticles ofSample 1-3 had the least dispersity. The dispersity difference betweenSamples 1-1 and 1-3 is clearly seen in FIG. 1 (Sample 1-1) and FIG. 2(Sample 1-3).

For a more accurate estimation of the nanoparticle size distribution ofsilver particle sizes versus bimetallic nanoparticles, dynamic lightscattering was applied. The average diameter of silver monometallicnanoparticles (Sample 1-1) was 9-10 nm (FIG. 4), while the bimetallicnanoparticles of Sample 1-3 were smaller at 5-6 nm (FIG. 5).

Example 3—Antimicrobial and Cytotoxic Properties—Bimetallic NanoparticleComposition with Miramistin®

Minimum inhibitory concentration (MIC) was determined to assessantibacterial and antifungal activities of the samples inmicroorganisms. MIC is the lowest concentration of a drug, whichprevents visible growth of bacterium. The tested panel of microorganismsrepresent a very broad spectrum of microorganisms, includingGram-positive and Gram-negative strains (Staphylococcus aureus vs.Escherichia coli), methicillin resistant strain INA 00761(Staphylococcus aureus), vancomycin resistant strain VKPM B-4177(Leuconostoc mesenteroides), fungi strain INA 00760 (Aspergillus niger),and yeast strain RIA259 (Saccharomyces cerevisiae). The results obtainedare presented in Table 2.

TABLE 2 Silver Nitrate Miramistin ® Sample 1-1 Sample 1-3 MicroorganismMIC, ug/mL Escherichia coli 10 20 1 0.5 ATCC 25922 Staphylococcus 20 5 51 aureus FDA 209P Leuconostoc 5 10 5 5 mesenteroides VKPMB-4177Staphylococcus 5 10 2.5 0.5 aureus INA 00761 Saccharomyces 10 20 5 2.5cerevisiae RIA 259 Aspergillus niger 20 20 5 0.5 INA 00760

As seen in the Table 2, the MIC of Sample 1-3 was significantly lowercompared to Miramistin®, silver nitrate, and a silver monometallicnanoparticle composition with Miramistin®. Which means that silver-goldbimetallic nanoparticles with Miramistin® (Sample 1-3) aresynergistically more effective against the microorganisms listedcompared to the controls.

The cytotoxic effects of Samples 1-1 and 1-3 on cancer cells werestudied in HCT-116 (human colon cancer cell lines), CRL-2945 (humanovarian carcinoma) and U-87 (human glioblastoma). Cell viability wasanalyzed in cells treated with different concentrations of Sample 1-1and Sample 1-3. Nanoparticle samples were able to reduce viability in adose-dependent manner. Viability was reduced significantly after 24 h oftreatment. The doses were used to calculate IC₅₀ value against thecontrol cells (Table 3).

TABLE 3 IC₅₀, ug/mL Cancer cell line Sample 1-1 Sample 1-3 HCT-116 20 10CRL-2945 20 10 U-87 15 5

As seen in the Table 3 the IC₅₀ of Sample 1-3 was lower compared tomonometallic silver nanoparticle composition with Miramistin® (Sample1-1). Thus, at least the silver-gold nanoparticle composition withMiramistin® of Sample 1-3 was more toxic to cancer cells compared tosilver monometallic nanoparticles with Miramistin®.

Example 4—Nanoparticle Preparation—Bimetallic Silver-Gold NanoparticleComposition with Miramistin® and Higher Gold Content

The silver-gold bimetallic nanoparticle compositions of Table 4 wereprepared according to the process of Example 1, wherein differentamounts of gold- and silver-bearing reagents were utilized. Themonometallic gold nanoparticle composition of Sample 2-4, lackingsilver, was prepared according to the process of Example 1, with theexception that silver salts were omitted.

TABLE 4 Sample No. Au molar % in Sample Sample 2-1 30 Sample 2-2 50Sample 2-3 70 Sample 2-4 100

Determination of the resulting nanoparticles size distribution wascarried out analogously to Example 2. The average diameter ofnanoparticles increased (15-20 nm). The dispersity of the colloids wasalso changed (data not shown).

Evaluation of antibacterial activity was carried out analogously toExample 3. The antibacterial activity of Samples 2-1 and 2-2 was higherthan Samples 2-3 and 2-4. Overall, all the samples of the presentexample revealed less antimicrobial activity compared to the samples ofExample 1 (see Table 5).

TABLE 5 Sample Sample Sample Sample 2-1 2-2 2-3 2-4 Microorganism MIC,ug/mL Escherichia coli 10 50 50 70 ATCC 25922 Staphylococcus 20 25 25 20aureus FDA 209P Leuconostoc 5 10 35 35 mesenteroides VKPMB-4177Staphylococcus 5 25 35 25 aureus INA 00761 Saccharomyces 10 20 35 70cerevisiae RIA 259 Aspergillus niger 20 25 25 20 INA 00760

Example 5—Nanoparticle Preparation—Bimetallic Silver-Gold NanoparticleComposition with Substitution of Miramistin®

Nanoparticle compositions were prepared analogously to Sample 1-3,wherein the surfactantbenzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) was substituted with cetyltrimethylammonium chloride(Sample 3-1), benzalkonium chloride (Sample 3-2),didecyldimethylammonium chloride (Sample 3-3), octenidinedihydrochloride (Sample 3-4), dimethyl benzyl ammonium chloride (Sample3-5), and polyhexamethylene biguanide chloride (Sample 3-6).

Determination of the resulting nanoparticles size distribution wascarried out analogously to Example 2. The average diameter ofnanoparticles ranged from 5 to 30 nm (data not shown).

Evaluation of antibacterial activity was carried out analogously toExample 3. The antibacterial activity of the resulting products exceededthe antibacterial activity of Sample 1-1 against a series of bacteria.

TABLE 6 Sample Sample Sample Sample Sample Sample 3-1 3-2 3-3 3-4 3-53-6 Microorganism MIC, ug/mL Escherichia — 1 — — 0.5 — coli ATCC 25922Staphylococcus 5   1 — 20   30 1   aureus FDA 209P Leuconostoc — — 5   —— — mesenteroides VKPMB-4177 Staphylococcus 5   1 — 2.5 — 0.5 aureus INA00761 Saccharomyces 2.5 — 2.5 — 10 2.5 cerevisiae RIA 259 Aspergillus2.5 — — — 5 — niger INA 00760

Example 6—Nanoparticle Preparation—Nanoparticle Composition withMiramistin® and Antibiotic

Nanoparticle compositions with silver, Miramistin®, and an antibioticwere prepared by method A or B. Method A: aqueous solution of a silversalt was added to a solution of at least one surfactant and antibioticunder vigorous stirring, then a reducing agent was added. The reactionwas carried out in an inert atmosphere of nitrogen or argon. Method B:aqueous solution of a silver salt was added to a solution of at leastone surfactant under vigorous stirring, then a reducing agent was added.Upon completion of the reaction at least one antibiotic was added, andthe resulting mixture stirred further. Silver nitrate or silver acetatecan be used as the silver salt in both methods.

Monometallic silver nanoparticle compositions with Miramistin® andtetracycline were prepared by Method A. In more detail, distilled waterwas repeatedly distilled in the atmosphere of nitrogen gas to achievede-oxygenation. The de-oxygenized water was used for all furtherpreparations. An aqueous solution of silver nitrate (20 mg in 10 mLwater) was added dropwise to vigorously stirred 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) and 100 uM tetracycline in water. The resulting mixturewas stirred for 15 min, followed by dropwise addition of 0.01 g sodiumborohydride in 100 mL water. After addition of all the reagents thereaction mixture was stirred for another hour. The preparation stepswere carried out in nitrogen or argon.

Bimetallic silver-gold nanoparticle compositions with Miramistin® andamoxicillin were prepared by Method A. In more detail, distilled waterwas repeatedly distilled in the atmosphere of nitrogen gas to achievede-oxygenation. The de-oxygenized water was used for all furtherpreparations. Solutions of 20 mg silver nitrate and 0.15 mg hydrogentetrachloroaurate in 10 mL water were added dropwise to vigorouslystirred 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) and 0.1 mg/mL amoxicillin in water. The resulting mixturewas stirred for 15 min, followed by dropwise addition of 0.01 g sodiumborohydride in 100 mL water. After addition of all the reagents thereaction mixture was stirred for another hour. The preparation stepswere carried out in nitrogen or argon.

Monometallic silver nanoparticle compositions with Miramistin andvancomycin were prepared by Method B. In more detail, 10 mL aqueoussolution of 20 mg silver nitrate was added dropwise to vigorouslystirred 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) in water. The resulting mixture was stirred for 15 min,followed by dropwise addition of 0.01 g sodium borohydride in 100 mLwater. After addition of all the reagents, the reaction mixture wasstirred for another hour. Then, 10 mL of 5 uM vancomycin water solutionwas added to the mixture and stirred for 10 h.

Table 7 shows the abbreviations used for obtained formulations.

TABLE 7 Sample No. Composition Sample 4-1 Ag NPs - Miramistin -tetracycline Sample 4-2 Ag—Au NPs - Miramistin - amoxicillin Sample 4-3Ag NPs - Miramistin- vancomycin

Example 7—Nanoparticle Characterization—Nanoparticle Composition withMiramistin® and Antibiotic

Nanoparticle compositions formed according to Example 6 werecharacterized. UV-V absorption spectra were measured to confirmnanoparticle formation. FIG. 7 shows the absorption spectra of theresulting compositions. All three compositions showed typical UVabsorption spectra for nanoparticles containing silver. Incorporation ofgold (Sample 4-2) affects the dispersity of the nanoparticles in thefinal product, and the absorption spectrum of the bimetallicnanoparticles displayed a more intense, narrow, and symmetricalabsorption band. TEM observations confirmed the formation ofnanoparticles in all 3 samples. Confirmation of association between thenanoparticles and antibiotics was done based on Raman spectra of thenanoparticles. Raman spectra of washed and dried Samples 4-1 through 4-3showed signals characteristic of tetracycline, amoxicillin and/orvancomycin thus demonstrating the association of the antibiotics withthe nanoparticles (data not shown).

For a more accurate estimation of size distribution of the silver andsilver-gold nanoparticles, dynamic light scattering was applied. Theresults obtained showed the average diameter of Sample 4-1 was 25 nm(FIG. 8A), Sample 4-2 was smaller at 19 nm (FIG. 8B), and Sample 4-3 was30 nm (FIG. 8C).

Example 8—Antimicrobial Properties—Nanoparticle Composition withMiramistin® and Antibiotic

Minimum inhibitory concentration (MIC) was determined to assess theantibacterial potential of the compositions of Example 6 inmicroorganisms. MIC is the lowest concentration of a drug which preventsvisible growth of bacterium. The tested panel of microorganismsrepresented a very broad spectrum of antibiotic resistantmicroorganisms. Sample 4-1 and Tetracycline (as control) were testedagainst multidrug-resistant Salmonella typhimurium DT 104. Sample 4-2and Amoxicillin (as control) were tested against Bacillus subtilis.Sample 4-3 and Vancomycin (as control) were tested against vancomycinresistant Enterococcus type Van-A and also vancomycin resistantLeuconostoc mesenteroides VKPM B-4177. In all test conditionsmonometallic silver nanoparticle composition with Miramistin® alone(obtained using a process known in the art) were used as a control. Thiscomposition is referred to as “Ag-NPs-Myramistin in Table 8. The resultsobtained are presented in Table 8.

TABLE 8 Sample Sample Sample Ag-NPs- 4-1 Tetracycline 4-2 Amoxicillin4-3 Vancomycin Myramistin Microorganism MIC, ug/mL Salmonella 4 >128 — —— — 10 typhimurium DT 104 Bacillus subtilis — — 15 >128 — — 30Leuconostoc — — — — 1 >128 5 mesenteroides VKPM B-4177 E. faecium A — —— — 2 >128 15 E. faecalis A — — — — 4 >128 15

As seen in Table 8, the MIC values of nanoparticle compositions with asurfactant and an antibiotic (Samples 4-1 through 4-3) are significantlylower compared to their corresponding antibiotics applied alone.Moreover, the antibiotics showed no efficacy against drug resistantpathogens. Although the silver nanoparticle preparation with Myramistinshowed antimicrobial activity, the nanoparticle compositions withMiramistin® showed a synergistic efficacy when combined with antibiotics(Samples 4-1, 4-2, and 4-3). The results demonstrate that silverMiramistin® coupled with surfactants and antibiotics not only cope withdrug-resistance, but also have very high antibacterial activity.

Example 9—Nanoparticle Preparation—Monometallic Silver NanoparticleComposition with Antibiotic and Substitution of Miramistin®

Nanoparticle compositions were prepared analogously as for Sample 4-1,wherein the surfactantbenzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) was substituted with cetyltrimethylammonium chloride(Sample 5-1), benzalkonium chloride (Sample 5-2),didecyldimethylammonium chloride (Sample 5-3), octenidinedihydrochloride (Sample 5-4), dimethyl benzyl ammonium chloride (Sample5-5), and polyhexamethylene biguanide chloride (Sample 5-6).

Determination of the resulting nanoparticle size distribution wascarried out analogously to Example 2. The average diameter of thenanoparticles ranged from 10 to 50 nm (data not shown).

Evaluation of antibacterial activity was carried out analogously toExample 3. The antibacterial activity of the resulting nanoparticlecompositions exceeded the antibacterial activity of silver colloidAg-NPs-Myramistin against a series of bacteria.

TABLE 9 Sample 5-1 Sample 5-2 Sample 5-3 Sample 5-4 Sample 5-5 Sample5-6 Microorganism MIC, ug/mL Escherichia coli 10 0.5 5 10 1 1 ATCC 25922Staphylococcus 20 5 10 — — 1 aureus FDA 209P Leuconostoc 10 10 20 5 10 5mesenteroides VKPM B-4177 Staphylococcus — — — 10 10 1 aureus INA 00761Saccharomyces — — — 5 2.5 5 cerevisiae RIA 259 Aspergillus niger 5 2.520 — — — INA 00760

Example 10—Nanoparticle Preparation—Monometallic Platinum NanoparticleComposition with Miramistin® and Glucosamine or Doxorubicin

Monometallic platinum nanoparticle compositions comprising Miramistin®and glucosamine or doxorubicin were prepared as follows: aqueoussolution of 50 mg hexachloroplatinic acid hexahydrate in 10 mL water wasadded dropwise to vigorously stirred aqueous 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®). The resulting mixture was stirred for 15 min, followed bydropwise addition of 0.01 g sodium borohydride in 100 mL water. Thecolor changed to dark brown. After addition of all the reagents, thereaction mixture was stirred for another hour. Then glucosamine (Sample6-1) or a doxorubicin water solution (Sample 6-2) was added, and theresulting mixture stirred further.

Table 10 shows the abbreviations used for the nanoparticle compositionsobtained.

TABLE 10 Sample No. Composition Sample 6-1 Pt NPs - Miramistin -Glucosamine Sample 6-2 Pt NPs - Miramistin - Doxorubicin

Example 11—Nanoparticle Preparation—Bimetallic Silver NanoparticleComposition with Miramistin® and Doxorubicin or Metformin

A bimetallic silver-platinum nanoparticle composition with Miramistin®and Metformin was prepared as follows: aqueous solution of silvernitrate (20 mg in 10 mL water) was added dropwise to vigorously stirred0.01 w/v % benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammoniumchloride (Miramistin®). The resulting mixture was stirred for 15 min,followed by dropwise addition of 100 mL aqueous 0.01 g sodiumborohydride. The color changed to brown. 2.5 mg hexachloroplatinic acidhexahydrate in 10 mL water was added dropwise to the resulting reactionmixture, stirred for 1 h, followed by dropwise addition of 10 mL aqueous1 mg sodium borohydride. After addition of all the reagents, thereaction mixture was stirred for another hour. Then a Metformin watersolution was added, and the resulting mixture stirred further.

A bimetallic silver-gold nanoparticle composition with Miramistin andDoxorubicin was prepared by Method A described in Example 6. In moredetail, distilled water was repeatedly distilled in the atmosphere ofnitrogen gas to achieve de-oxygenation. The de-oxygenized water was usedfor all further preparations. Aqueous solutions of 20 mg silver nitrateand 0.15 mg hydrogen tetrachloroaurate were added dropwise to vigorouslystirred 150 mL 0.01 w/v %benzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin) in water. The resulting mixture was stirred for 15 min,followed by dropwise addition of 0.01 g sodium borohydride in 100 mLwater. After addition of all the reagents the reaction mixture wasstirred for another hour. Then a Doxorubicin water solution was added,and the resulting mixture stirred further. Table 11 shows theabbreviations used for the obtained compositions.

TABLE 11 Sample No. Composition Sample 7-1 Pt—Ag NPs - Miramistin -Metformin Sample 7-2 Au—Ag NPs - Miramistin - Doxorubicin

Example 12—Nanoparticle Characterization—Platinum Nanoparticles

UV-V is absorption spectra were measured to confirm formation of thenanoparticles in the compositions of Examples 10 and 11. FIG. 9 showsthe absorption spectra of the resulting nanoparticles. All 4nanoparticles showed UV absorption spectra typical for platinumnanoparticles. Incorporation of silver (Sample 7-1) resulted in a redshift in the UV spectrum. TEM observations confirmed the formation ofnanoparticles in all 4 samples (FIGS. 10A-10C). The confirmation ofdoxorubicin associated with the nanoparticles was done based on Ramanspectra. Raman spectra of washed and dried Samples 6-2 and 7-2 showedsignals characteristic of doxorubicin (data not shown). For a moreaccurate estimation of the nanoparticle size distribution, dynamic lightscattering was applied. The average diameter of Sample 6-1 was 4.5 nm(FIG. 11A), Sample 6-2 was slightly smaller at 4 nm (FIG. 11B), Sample7-1 was 3 times bigger at 13 nm (FIG. 11C), and Sample 7-2 was around 5nm (data not shown).

Example 13—Anticancer Properties—Platinum and Silver ContainingNanoparticle Compositions with Miramistin® and Drug

The cytotoxic effects of Samples 6-1, 6-2, 7-1 and 7-2, glucosamine andMetformin on cancer cells were studied in CRL-2945 (human ovariancarcinoma) and U-87 (human glioblastoma). The doxorubicin resistantCRL-2945 and U-87 cells were generated by exposing the cells to thecytotoxic drug at incrementally increasing concentrations. In this way,resistant sublines were obtained (CRL-2945-R and U-87-R). The cellviability was analyzed in cells treated with different concentrations ofSample 6-1, Sample 6-2, Sample 7-1 and Sample 7-2. The samples were ableto reduce viability in a dose-dependent manner. Viability reducedsignificantly after 24 h of treatment. The doses were used to calculateIC₅₀ values against the control cells (Table 12).

TABLE 12 IC₅₀ Sample, Sample Sample Sample Cancer Doxorubicin, 6-1 6-2,7-1, 7-2, cell line uM ug/mL ug/mL ug/mL ug/mL CRL- 0.13 40 5 20 25 2945U-87 0.08 20 2 10 20 CRL- 0.28 40 7 20 30 2945 -R U-87 -R 1.2 15 8 10 20

As seen in the Table 12 the IC₅₀ of Sample 6-2 is the lowest compared tothe other compositions tested. Higher toxicity of Sample 6-2 can beexplained by the fact that Sample 6-2 bears very toxic doxorubicin,which is delivered into the cells with nanoparticles. Doxorubicinresistant cell sublines were significantly less resistant tonanoparticles with doxorubicin (i.e. Sample 6-2). Sample 6-1 and Sample7-1 showed high cell toxicity at higher concentrations, but affected theamount of cancer stem cells (CSCs) as seen in Table 13. Cancer cellsCRL-2945-R and U-87-R were cultured in serum-free media to enrich thepopulations with cancer stem cells. Thus, enriched cells were testedwith the samples. Glucosamine and Metformin alone did not show anysignificant activity (data not shown).

TABLE 13 Before treatment After treatment with After treatment with withNPs Sample 6-1, 40 ug/mL Sample 7-1, 20 ug/mL Cancer CD44, % CD24, %CD44, % CD24, % CD44, % CD24, % cell line positive cells positive cellspositive cells positive cells positive cells positive cells CRL-2945 2023 12 50 7 58 U-87 55 20 42 45 33 56

As seen in the Table 13 the percentage of CSC (CD44 high, CD24 low)decreased after treatment of the cells with Sample 6-1 and 7-1. Sample7-1 had a more prominent effect.

Example 14—Nanoparticle Preparation—Monometallic Platinum NanoparticleComposition with Doxorubicin and Substitution of Miramistin®

Nanoparticle compositions were prepared analogously to Sample 6-2,wherein the surfactantbenzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride(Miramistin®) was substituted with cetyltrimethylammonium chloride(Sample 8-1), benzalkonium chloride (Sample 8-2),didecyldimethylammonium chloride (Sample 8-3), octenidinedihydrochloride (Sample 8-4), dimethyl benzyl ammonium chloride (Sample8-5), and polyhexamethylene biguanide chloride (Sample 8-6).

Determination of the resulting nanoparticle size distribution wascarried out analogously to Example 12. The average diameter ofnanoparticles ranged from 5 to 45 nm (data not shown).

Evaluation of anticancer activity was carried out analogously to Example13. The anticancer activity of the resulting products is also high andsimilar concentration ranges.

TABLE 14 Cancer IC₅₀ cell line Sample 8-1 Sample 8-2 Sample 8-3 Sample8-4 Sample 8-5 Sample 8-6 CRL-2945 25 15 20 20 20 10 U-87 10 20 20 15 155 CRL-2945-R 20 10 25 20 25 10 U-87-R 10 15 20 20 10 5

Table 15 presents the various embodiments of the nanoparticlesexemplified above, including their corresponding examples.

TABLE 15 Relevant Sample No. Composition Examples Sample 1-1Monometallic silver nanoparticles with Examples 1, Miramistin ® 2, 3,and 5 Sample 1-2 Bimetallic silver-gold nanoparticles Examples 1 withMiramistin ®; 99.67% molar Ag to and 2 0.33% molar Au Sample 1-3Bimetallic silver-gold nanoparticles Examples 1, with Miramistin ®;99.50% molar Ag to 2, 3, and 4 0.50% molar Au Sample 1-4 Bimetallicsilver-gold nanoparticles Example 1 with Miramistin ®; 97.50% molar Agto 2.50% molar Au Sample 2-1 Bimetallic silver-gold nanoparticlesExample 4 with Miramistin ®; 70% molar Ag to 30% molar Au Sample 2-2Bimetallic silver-gold nanoparticles Example 4 with Miramistin ®; 50%molar Ag to 50% molar Au Sample 2-3 Bimetallic silver-gold nanoparticlesExample 4 with Miramistin ®; 30% molar Ag to 70% molar Au Sample 2-4Monometallic gold nanoparticles with Example 4 Miramistin ® Sample 3-1Bimetallic silver-gold nanoparticles Example 5 withcetyltrimethylammonium chloride; 99.50% molar Ag to 0.50% molar AuSample 3-2 Bimetallic silver-gold nanoparticles Example 5 withbenzalkonium chloride; 99.50% molar Ag to 0.50% molar Au Sample 3-3Bimetallic silver-gold nanoparticles Example 5 withdidecyldimethylammonium chloride; 99.50% molar Ag to 0.50% molar AuSample 3-4 Bimetallic silver-gold nanoparticles Example 5 withoctenidine dihydrochloride; 99.50% molar Ag to 0.50% molar Au Sample 3-5Bimetallic silver-gold nanoparticles Example 5 with dimethyl benzylammonium chloride; 99.50% molar Ag to 0.50% molar Au Sample 3-6Bimetallic silver-gold nanoparticles Example 5 with polyhexamethylenebiguanide chloride; 99.50% molar Ag to 0.50% molar Au Sample 4-1Monometallic silver nanoparticles with Examples 6, Miramistin ® andtetracycline 7, and 8 Sample 4-2 Bimetallic silver-gold nanoparticlesExamples 6, with Miramistin ® and amoxicillin 7, and 8 Sample 4-3Monometallic silver nanoparticles with Examples 6, Miramistin ® andvancomycin 7, and 8 Sample 5-1 Monometallic silver nanoparticles withExample 9 cetyltrimethylammonium chloride and tetracycline Sample 5-2Monometallic silver nanoparticles with Example 9 benzalkonium chlorideand tetracycline Sample 5-3 Monometallic silver nanoparticles withExample 9 didecyldimethylammonium chloride and tetracycline Sample 5-4Monometallic silver nanoparticles with Example 9 octenidinedihydrochloride and tetracycline Sample 5-5 Monometallic silvernanoparticles with Example 9 dimethyl benzyl ammonium chloride andtetracycline Sample 5-6 Monometallic silver nanoparticles with Example 9polyhexamethylene biguanide chloride and tetracycline Sample 6-1Monometallic platinum nanoparticles Examples 10, with Miramistin ® andglucosamine 12, and 13 Sample 6-2 Monometallic platinum nanoparticlesExamples 10, with Miramistin ® and doxorubicin 12, and 13 Sample 7-1Bimetallic silver-platinum nanoparticles Examples 11, with Miramistin ®and metformin 12, and 13 Sample 7-2 Bimetallic silver-gold nanoparticlesExamples 11, with Miramistin ® and doxorubicin 12, and 13 Sample 8-1Monometallic platinum nanoparticles Example 14 withcetyltrimethylammonium chloride and glucosamine Sample 8-2 Monometallicplatinum nanoparticles Example 14 with benzalkonium chloride andglucosamine Sample 8-3 Monometallic platinum nanoparticles Example 14with didecyldimethylammonium chloride and glucosamine Sample 8-4Monometallic platinum nanoparticles Example 14 with octenidinedihydrochloride and glucosamine Sample 8-5 Monometallic platinumnanoparticles Example 14 with dimethyl benzyl ammonium chloride andglucosamine Sample 8-6 Monometallic platinum nanoparticles Example 14with polyhexamethylene biguanide chloride and glucosamine

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1-39. (canceled)
 40. A method of treating cancer comprisingadministering to a patient in need thereof an effective amount of acomposition comprising (i) particles having at least one metal, and (ii)at least one surfactant.
 41. The method of claim 40, wherein said atleast one metal comprises a metal selected from silver, gold, platinum,palladium, osmium, iridium, rhodium, ruthenium, and combinationsthereof.
 42. The method of claim 40, wherein said particles includemonometallic particles, polymetallic particles, or a combinationthereof.
 43. The method of claim 42, wherein said polymetallic particlesinclude bimetallic particles.
 44. The method of claim 43, wherein saidbimetallic particles include silver and gold bimetallic particles,silver and platinum bimetallic particles, or a combination thereof. 45.The method of claim 43, wherein said bimetallic particles have a corethat includes at least one of gold and platinum, and a shell thatincludes silver.
 46. The method of claim 42, wherein said monometallicparticles are selected from silver monometallic particles, goldmonometallic particles, platinum monometallic particles, andcombinations thereof.
 47. The method of claim 40, wherein said particlesinclude nanoparticles.
 48. The method of claim 40, wherein said at leastone surfactant comprises a surfactant selected from a cationicsurfactant, an anionic surfactant, an amphoteric surfactant, acharge-neutral surfactant, and combinations thereof.
 49. The method ofclaim 40, wherein said at least one surfactant is selected frombenzyl-dimethyl-[3-(tetradecanoylamino)propyl]ammonium chloride, acetyltrimethylammonium salt, a benzalkonium salt,didecyldimethylammonium chloride, octenidine dihydrochloride, dimethylbenzyl ammonium chloride, a polyhexamethylene biguanide, apolyhexamethylene guanidine, a polyhexamethylene biguanide salt, apolyhexamethylene guanidine salt, sodium lauryl ether sulfate, sodiumcocaminopropionate, and combinations thereof.
 50. The method of claim40, wherein said composition further comprises a drug.
 51. The method ofclaim 50, wherein said drug is selected from doxorubicin, glucosamine,metformin, and combinations thereof.
 52. The method of claim 40, whereinsaid cancer is selected from glioblastoma, colon cancer, and ovariancancer.
 53. The method of claim 40, wherein said composition furthercomprises an antibiotic.
 54. The method of claim 53, wherein saidantibiotic is selected from tetracycline, vancomycin, ampicillin,amoxicillin, penicillin, methicillin, enoxacin, ofloxacin, norfloxacin,ciprofloxacin, kanamycin A, amikacin, tobramycin, teicoplanin,telavancin, and combinations thereof.
 55. The method of claim 40,wherein said composition further comprises at least one of a carrier andan excipient.
 56. The method of claim 40, wherein said composition is acolloid.
 57. The method of claim 40, wherein said composition isadministered in one or more forms selected from a pill, capsule, tablet,microbead, injection, infusion, and suppository.